AIR-O-CELL ®

Bioaerosol Sampling Cassette

Laboratory
& User Manual

LA03040 Rev.2
 Air-O-Cell® Sampling Cassette

The Air-O-Cell is a unique sampling cassette specifically designed for the rapid collection
and quantitative analysis of a wide range of airborne aerosols. It collects both viable and
non-viable particulate such as mold spores, pollen, insect parts, skin cell fragments, fibers
(asbestos, fiberglass, cellulose, etc.) and inorganic particles.

Suggested & potential applications include but are not limited to the following:

APPLICATIONS
Indoor Air Quality: Mold spores, pollen, insect parts, dust mites, skin cell frag-
ments, plant fragments, dust, fibers, combustion emissions, etc.
Home Inspection: Mold Contamination before of after real estate transactions.
Flood Restoration: Evaluation of mold spore contamination before, during, and
after remediation.
Allergy Testing: Mold spores, pollen, insect parts, dust mites.
Clean Room Monitoring: Evaluation of low airborne dust and contaminants from
personnel (skin cells, clothing fibers, cosmetics, etc.)
Fiber Analysis: Asbestos, fiberglass, cellulose, ceramics, etc.
Stack Emissions: Fly ash, inorganic dust, etc.

Air-O-Cell® ADVANTAGES

• Provides excellent detection limits over conventional filter sampling utilizing

25 mm or 37 mm diameter filter cassettes.
• Eliminates sample loss to cassette walls known to occur with filter samples
from vibration or static charge during sampling and shipment.
• Eliminates the need for direct handling or preparation of collection media or
microscope slides in the field.
• Eliminates potential cross-contamination between samples and during ship-
ping that may occur with other devices.
• Unique optically transparent and smooth collection media allows direct stain-
ing and examination by bright field, dark field, and phase contrast microscopy!!
• The sampling media is compatible with a wide range of biological stains and
refractive index oils allowing for direct quantitative analysis of biological and
inorganic particles.
• The Air-O-Cell will work with virtually any kind of sampling pump
capable of pulling a 15 lpm (vacuum) air flow.

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 PRINCIPLE OF OPERATION

The Air- O-Cell operates on the well established principle of iner tial
impaction. Particles in the air stream are accelerated as they approach the tapered inlet
opening and drawn through a small slit aimed directly at a glass slide. This glass slide
contains a sticky and optically clear sampling media which can permanently collect and
hold particles. As the particles come through the slit, the air velocity forces the particles
to impact into the sampling media, while the air stream makes a sharp 90o turn and
proceeds around the slide and out of the cassette. The air flow path through the
Air-O-Cell cassette is illustrated below in Figure 1.

Figure 1 AIR FLOW PATH OF THE Air-O-Cell

air flow into cassette

sample slit

collection media

glass slide

exit orifice

lateral view

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 RECOMMENDED SAMPLING PROCEDURES

General:

The Air-O-Cell sampler is designed to operate at an optimal flow rate of 15 liters per
minute. The user can employ any sampling pump capable of a minimum flow rate
of 15 lpm. It is also capable of operating in any vertical or horizontal orientation, or
in restricted access spaces smaller than 2 inches in diameter. As a result the Air-
O-Cell is ideally suited for sampling in HVAC ducts, plenums, wall cavities, or other
confined spaces.

Sampling of Ambient Static Environments:

A rotameter calibrated to a primary standard, soap bubble tube/meter or a dry

bubble meter should be used to calibrate the sampling pump to a flow rate of
15 lpm. Some pumps only work with specific calibration devices. Please reference
the owners manual for your pump to verify if any special calibration methods should
be employed. Because the cassette does not produce significantly measurable back
pressure, the rotameter can optionally be connected directly to the pump (without
the Air-O-Cell cassette in line) to calibrate the pump flow rate.

To begin sampling, remove the tape seals covering the inlet and outlet and placed
them on the side of the cassette. Then connect the Air-O-Cell cassette to the sam-
pling pump using flexible tubing. Turn the sampling pump on for an appropriate
sample time ranging from 1 to 10 minutes and both seals replaced after sampling is
complete. Unlike other spore trap impaction or filter devices, the Air-O-Cell cassette
can be oriented in any vertical or horizontal direction, without concern for sample
loss of large particles or vibration. “Outdoor background” samples should always be
collected for comparison purposes.

Sampling in HVAC Systems:

The Air-O-Cell cassette design allows for isokinetic sampling of aerosols in Heating,
Ventilation and Air Conditioning (HVAC) Systems. Sampling can be conducted at the
supply diffuser or inside most conventional ducts. The inlet of the cassette should
always be facing into the flow stream. The inlet orifice has a cross-sectional area
of approximately 11 mm x 15 mm (165.0 mm2) tapering to a slit with dimensions
of 1.055 mm x 14.4 mm (15.19 mm2). The flow velocity can be increased up to
30 lpm with conventional sampling pumps, however, air flows exceeding 20 lpm
may potentially damage some bioaerosols or cause “bounce off”. Isokinetic sampling
can be conducted in most air duct system with flow rates of up to approximately
600 fpm. Approximate face velocities for the Air-O-Cell cassette are given below for
both the entrance orifice and slit exit in Table 1

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Table 1
Air-O-Cell Theoretical Face Velocities

Flow Rate Orifice Face Orifice Face Slit Face Slit Face
(lpm) Velocity Velocity Velocity Velocity
(fpm) (mph) (fpm) (mph)

15.0 299 3.4 3110 35.3

20.0 399 4.5 4146 47.2
25.0 499 5.7 5183 59.0
28.3 (1 cubic ft.) 564 6.4 5867 66.8
30.0 598 6.8 6219 70.6

Recommended Sampling Time Intervals:

Although the Air-O-Cell cassette can provide excellent detection limits over con-
ventional filter sampling utilizing 25 mm or 37 mm diameter filter cassettes, it is
also sensitive to overloading. In an appropriately loaded sample, the trace should
be barely visible and transparent, but not opaque or dense. If the sample appears
highly visible or opaque, additional shorter time interval samples should be collected.
The recommended sampling flow rate is 15 liters per minute (lpm). As mentioned
above, flow rates exceeding 20 lpm have been known to cause “bounce off” of
large particles such as pollen grains. Flow rates lower than 10 lpm will not collect
the small mold spores (such as Aspergillus and Penicillium) as efficiently. Recom-
mended sampling times (at 15 lpm) for different environmental sampling conditions
are given in Table 2.

Table 2

RECOMMENDED SAMPLING INTERVALS

Sampling Time (minutes)

Environmental Dust Conditions 15 lpm

• Outdoor sampling on a clean windless day 10.0 - 60 min.

• “Clean” office environment or outdoors (no visible dust) 10.0 min.
• “Indoor” environment, high activity personnel 5.0 min.
• “Indoor” environment, evidence of drywall renovation, or
industrial dust 1.0 min.
• “Indoor” environment, visible dust emissions from point
sources present 0.5 min.

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 RECOMMENDED LABORATORY ANALYSIS
PROCEDURES

Slide preparation

One to two (1-2) drops of staining or mounting media (lacto-phenol cotton blue is
recommended for mold spore analysis) should be placed in the center of a clean
pre-labeled slide. Air-O-Cell cassettes should only be opened in the laboratory.
The sealing band should be cut, and the glass cover slip (containing the sample
trace) removed and slowly placed on an angle with the media collection side down
onto the staining solution. Do not press down on the slide during or after staining!.
Excess staining solution should be removed from around the edges of the cover slip
with a tissue wipe or cotton swab after 10 minutes has elapsed. This will ensure
even staining of the sample. It should be noted that the slide can also optionally
be mounted media side up. To do this, use a drop of fingernail polish to secure the
Air-O-Cell slide to the microscope slide. Then place a couple drops of stain on the
media and place a cover slip on top.

About Stains

Numerous stains may be used during laboratory analysis. These include

lacto-phenol cotton blue, anline blue, calbreras stain and acid fusion
stain. The most common stain used for mold spore analysis is lacto-
phenol cotton blue.

To achieve the best clarity of the sample, using stains that have little or
no water content is preferred. Water can cause the sample to appear
cloudy.

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 Microscopic Examination
Analysis of the collected sample should be performed by an experienced Microbiolo-
gist, Aerobiologist, Mycologist or Environmental Microscopist. Counting and quanti-
fication of sample components is conducted by counting calibrated cross-sections
of the deposited sample trace. The number and type of particles counted per cubic
meter of air is calculated based on the length of the deposition trace, length of trace
actually examined, volume of air collected, and number of particles counted.

The Air-O-Cell particle deposition area at a flow rate of 15 lpm is approximately

1.1 mm wide by 14.5 mm long yielding an approximate area of 15.95 mm2. The
width of the deposition trace will vary slightly with flow rate and media thickness,
and will vary slightly in particle density from the middle to outer edges of deposi-
tion. For this reason, using the deposition trace area is not recommended for direct
calculation of particle concentrations. The recommended procedure for calculating
particle concentrations is based on using the Air-O-Cell trace length and microscope
field diameter, and will be discussed below. One field of view counted is defined as
the calibrated diameter of the microscope field of view (in mm) covering one cross-
sectional pass or “traverse” across the sample deposition trace. A typical sample
preparation and microscopic counting procedure is illustrated in Figure 2.

Figure 2
ILLUSTRATION OF Air-O-Cell COUNTING METHOD

microscopic slide

Air-0-Cell™ slide
analysis starts at 1st
traverse - completes and
moves to 2nd traverse

sample particulate

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The calculation of particle concentration per cubic meter of air can be performed
by using the following equations.

First, determine the actual air volume collected in cubic meters (m3) by following
the calculation given in Equation 1.

EQUATION 1:

Air volume (m3) = (Sampling rate (liters per minute) / 1000) x

Number of minutes

Second, determine the length of sample trace counted based on the microscope field
of view and number of fields of view counted. Accurately calibrate and measure the
diameter of the microscope field of view using a stage micrometer slide. Remember,
each microscope is different, and each different combination of ocular and objec-
tive lens must be calibrated separately. Stated lens magnifications are rarely precise.
The microscopist should then record the number of complete traverses examined
across the width of the deposition trace and use the formula given in Equation 2 to
calculate the actual length of the deposition trace examined.

EQUATION 2:

Trace Length Counted (mm2) = Microscope field

diameter (mm) x number of traverses

The concentrations of particles (cts/m3) can then be determined by using

Equation 3.

EQUATION 3:

Trace length (14.4 mm) 1

Cts / m3 = _______________ x ___________ x # of particle counts
Total length of Air Volume(m3)
trace counted
(From Equation 2) (From Equation 1)

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Two example calculations for mold spores and pollen grains are given below:

Example 1 Mold Spore Example

Microscope field diameter at (900X) = 0.240 mm

Number of traverses = 10
Sample volume (15 lpm @10 minutes) = (15 / 1000) x 10 = 0.150 m3
Mold spore counts = 50

14.4 mm 1 14.4
_________ x _______ x 50 = _____ x 50 = 2000 ct/m3
0.240 x 10 0.15 0.36

Example 2 Pollen Example

Microscope field diameter at (200X) = 1.11 mm

Number of traverses = 10
Sample volume (15 lpm @ 30 minutes) = (15 / 1000) x 30 = 0.450 m3
Pollen counts = 25
14.4 mm 1 14.4
________ x ______ x 25 = _____ x 25 = 72 grains/m3
1.11 x 10 0.45 5.0

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 RECOMMENDED MICROSCOPIC
COUNTING GUIDELINES
Counting & Identification Guidelines

Pollen — Entire trace or 100 grains (whichever comes first) should

be examined at a minimum magnification of 200X. Iden-
tification and speciation should be performed at minimum
magnification of 400X.

Mold Spores — A minimum of 15% of the entire trace should

be examined or a minimum of 100 mold spores
counted (whichever comes first). Identification and
s p e c i a t i o n s h o u l d b e p e r fo r m e d a t m i n i m u m
magnification of 400X.

Fibers — The entire trace or 100 fibers, (whichever comes first)

should be examined at a minimum magnification of 200X.

Other Aerosols — Skin cell fragments, combustion emissions, insect parts —A

minimum of 10% of the entire trace should be examined
or a minimum of 100 particles counted (whichever comes
first).

Storage & Operating Conditions

This product should be stored at room temperature, between 60-82°F (15-28°C).

Do not use product at temperatures below 32°F (0°C). If product has been
exposed to freezing temperatures immediately before sampling, it is recom-
mended to let the product acclimate to the sampling environment before use.

All bioaerosol samplers are not created equal and will not
provide the same results. Look for this seal on your product
as assurance of validated, industry proven performance and
the highest quality product.

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 Ordering Information
Field Equipment
Product Number Description
AOC010 Air-O-Cell® Cassettes, 10/box
AOC050 Air-O-Cell® Cassettes, 50/box
AOCWS10 Inner Wall Sampling Adapter, 10/box
AOC-CAL In-Line Calibration Adapter
ZBP-307 Zefon Bio-Pump® Bubble Tube for Air-O-Cell
ZBP-200 Zefon Bio-Pump®
APB-803300 mini-Buck IAQ Calibrator
DCL-H Bios DryCal® DC-Lite Calibrator

Laboratory Equipment
Product Number Description
ZA0046 Cassette Opener
ZA0050 Particle Spreader, “T” shaped, sterile
ZA0051 Syringe, sterile, 3cc
ZA0052 Alcohol wipes, 200/bx
ZA0054 Forceps, self closing, each

Storage & Operating Conditions

This product should be stored at room temperature, between 60-82°F (15-28°C).

Do not use product at temperatures below 32°F (0°C). If product has been
exposed to freezing temperatures immediately before sampling, it is recom-
mended to let the product acclimate to the sampling environment before use.

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 5350 SW 1st Lane, Ocala, FL 34474 • www.zefon.com
Phone: 800-282-0073 • 352-854-8080 • Fax: Fax: 352-854-7480
© Copyright 2009 Zefon International, Inc.
Via-Cell, Air-O-Cell, and BioPump are registered trademarks of Zefon International.

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